Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 1

herPetozoA 28 (1/2): 15 - 27 15 Wien, 30. Juli 2015

Classification of the mountain of the Berçelan Plateau (hakkari), east () (Anura: ranidae)

systematische zuordnung der Braunfrösche des Berçelan Plateaus (hakkari), ost-Anatolien (türkei) (Anura: ranidae)

murAt AfsAr & B irgül AfsAr & h üseyin ArikAn

kurzfAssung die Braunfrösche des Berçelan Plateaus (karadağ, hakkari) in ost-Anatolien wurden im hinblick auf ihre äußere morphologie (morphometrisch und nach färbungsmerkmalen) sowie serologisch (Plasma-Proteinelektro- phorese) untersucht. danach und im vergleich mit material der typischen fundorte von Rana macrocnemis Boulenger , 1885 und Rana holtzi Werner , 1898 zeigten die frösche des Berçelan Plateaus große übereinstim- mung mit den tieren des uludağ (Bursa), die derzeit R. macrocnemis zugeordnet werden.

ABstrACt in the present study, the mountain population of the Berçelan Plateau (karadağ, hakkari) in east Anatolia was examined both in terms of their morphology (morphometric and color-pattern characteristics) and serology (blood plasma protein electrophoresis). According to these studies and when compared with frogs from the type localities of Rana macrocnemis Boulenger , 1885, and Rana holtzi Werner , 1898, the mountain frogs of the Berçelan Plateau strongly resembled the specimens from uludağ (Bursa) which are currently assigned to R. macrocnemis . key Words Amphibia: Anura: ranidae; Rana macrocnemis , , morphology, polyacrylamide gel disc elec - trophoresis; Berçelan Plateau, east Anatolia, turkey

introduCtion

Although Rana macrocnemis Bou- ic and considered the taxa tavasensis BArAn lenger , 1885 , Rana holtzi Werner , 1898, & A tAtür , 1986, and pseudodalmatina and Rana camerani Boulenger , 1886, were eiselt & s Chmidtler , 1971 , as separate initially described as three separate , species, which had been classified as sub - different views about their systematic posi - species of R. macrocnemis . A recent mor - tions were set forth in the following years phological, morphometric and serological (Boulenger 1898; W erner 1902; l Antz & study on Anatolian mountain frogs by Cyrén 1913; B odenheimer 1944; m ertens Çevik et al. (2006) found two species ( R. 1952; B Aşoğlu & h elmiCh 1958, e iselt camerani and R. holtzi ) living in Anatolia 1965; B ArAn 1969; B ArAn & A tAtür , and considered R. camerani from mt. 1986; P iCAriello et al. 1999; tArkhni- erciyes a synonym of R. macrocnemis . shvili et al. 1999; ArikAn et al. 2001; subsequently, the mountain frog spec - düşen et al. 2002; BArAn et al. 2004). imens collected from new localities in Based on studies at the molecular Anatolia were examined in terms of their level, veith et al. (2003a, 2003b) regarded morphological characteristics and classified the Anatolian frog species R. m. macrocne - according to their taxonomic position, as mis , R. camerani and R. holtzi as conspecif - accepted at that time ( BArAn et al. 2007; Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 2

16 m. A fsAr & B. A fsAr & h. A rikAn

BAşkAle et al. 2012; AfsAr et al. 2013). in summary, R. holtzi was detected at two localities in the Bolkar mountains and near eğrigöl (Çamlıyayla, mersin) ( BArAn et al. 2007) and karagöl (Çamlıyayla, mersin) (BAşkAle et al. 2012) in the central , where its range appeared to be wider than expected. new localities of R. macrocnemis were determined from konya (ereğli, seviçova) and elazığ (maden, Örtülü village) by BArAn et al. (2007), and the southernmost record of the species was detected at the village of Örtülü (maden/ elazığ). however, the taxonomic position of the mountain frog populations in the hakkari section of the east Anatolian region remained unclear due to insufficient materi - al (see BArAn & A tAtür 1986). in this early paper, the populations of muş, and erciş were included in R. macrocnemis , whereas a specimen from mt. nemrut (Bitlis) was assigned to R. camerani . in the present study, numerous moun - tain frog specimens collected from the Berçelan Plateau (karadağ, hakkari) were examined with regard to their morphology (morphometric and color-pattern character - istics) and serology, with the aim to clarify the taxonomic position of the mountain frogs in the east Anatolian region.

mAteriAls And methods

study area and individuals studied.- the mountain frog specimens examined were obtained from the Berçelan Plateau, located at an altitude of 2,700 m on mt. karadağ in the north of hakkari (fig. 1). the frogs were collected on the moist

______

fig. 1: distribution of Rana macrocnemis Boulenger , 1885 , Rana tavasensis BArAn & A tAtür , 1986, and Rana holtzi Werner , 1898, in turkey. data according to BArAn & A tAtür (1988), veith et al. (2003), BArAn et al. (2007) and BAşkAle et al. (2012) green dots - R. macrocnemis , T - R. tavasensis , H - R. holtzi (Bolkar mountains) and red dot marked with BP - the localities of the Berçelan Plateau (hakkari) from which the specimens of the present study were collected . Abb. 1: verbreitung von Rana macrocnemis Boulenger , 1885 , Rana tavasensis BArAn & A tAtür , 1986 und Rana holtzi Werner , 1898 in der türkei nach Angaben in BArAn & A tAtür (1988), veith et al. (2003), BArAn et al. (2007) und BAşkAle et al. (2012). grüne Punkte - R. macrocnemis , T - R. tavasensis , H - R. holtzi (Bolkar gebirge), mit BP gekennzeichneter roter Punkt – die fundorte im Berçelan Plateau (hakkari), woher die in der vorliegenden Arbeit untersuchten frösche stammen. Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 3

the mountain frogs of the Berçelan Plateau (hakkari), east Anatolia 17

fig. 2: of Rana macrocnemis Boulenger , 1885, in the Berçelan Plateau (hakkari). Abb. 2: lebensraum von Rana macrocnemis Boulenger , 1885 im Berçelan Plateau (hakkari).

soil of the meadowland along the banks of mately be incorporated into the collection of small streams generated by the melting the zoology museum of ege university snow in open fields, and by spring waters (zdeu). Comparative data were obtained (fig. 2). from specimens of the type localities of R. the color-pattern characteristics of macrocnemis (zdeu 162/2002-1-12, f, m, those specimens were noted, which were uludağ, AfsAr & A fsAr leg. 17.07.2002) brought alive to the laboratory, and color and Rana holtzi (zdeu 141/1970-1-18 f, m, photographs taken. the specimens were karagöl, leg. BArAn 08.06.1970). preserved at the department of zoology at morphometric characters and Celal Bayar university (CBu, manisa). m – ratios.- the following morphometric males, f – females. measurements were taken: snout-vent materials.- CBu 9/2011, 1-12 m, length (svl), head length (hl), head width f, Berçelan, hakkari, 18.08.2011, leg. m. (hW), internarial distance (id), interorbital AfsAr & B. A fsAr , 37°39.407’n, 43°46. distance (io), femur length (fl), tibia 560’e. CBu 9/2011, 1-42 m, f, Berçelan, length (tl), hind leg length (AAu) and hakkari, 17.07.2012, leg. m. A fsAr & B. metatarsal tubercle length (CAl) were AfsAr , 37°39.555’n, 43°46.341’e. CBu measured using a compass sensitive to 0.01 1/2013, 1-8 m, f, uludağ, Bursa, 10.07. mm, according to terentJev & C hernoW 2013, leg. m. A fsAr & B. A fsAr , 40°07. (1965) and BArAn (1969). the ratios hl/ 905’n, 29°06.733’e (the 8 uludağ speci - svl, hW/svl, id/svl, io/svl, fl/svl, mens were used for blood plasma electro- tl/svl, AAu/svl and CAl/svl were phoresis only). the specimens will ulti - calculated from the morphological data. Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 4

18 m. A fsAr & B. A fsAr & h. A rikAn

figs. 3a-3d: four types of dorsal pattern found in Rana macrocnemis Boulenger , 1885, from the Berçelan Plateau (hakkari). Abb. 3a-3d: die vier typen der rückenfleckung wie sie bei Rana macrocnemis Boulenger , 1885 vom Berçelan Plateau (hakkari) gefunden wurden.

statistical analysis.- the data the help of the presence/absence of a callus under study was normally distributed ac - on the thumb as an indicator. the morpho - cording to the kolmogorov-smirnov test ( p metric data of males and females was com - > 0.05). Adult specimens were sexed with pared by using the independent samples t- Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.201514:54seite5

table 1: descriptive statistics of the morphometric data [mm] and ratios of the examined mountain frog populations from the Berçelan Plateau (hakkari), uludağ (Bursa) and karagöl, Bolkar mountains (niğde). n - number of specimens, m - mean; sd - standard deviation; se - standard errors of the mean. see materials and methods for abbreviations in column one. t

tab 1: deskriptive statistiken der morphometrischen daten (in mm) und Quotienten der untersuchten Braunfrosch-Populationen vom Berçelan Plateau (hakkari), h e uludağ (Bursa) und von karagöl, Bolkar gebirge (niğde). n – Anzahl exemplare, m – mittelwert; sd – standardabweichung, se – standardfehler des mittelwertes. m

die Abkürzungen in spalte eins sind im Abschnitt materials and methods erklärt. o u n t a

Characters Berçelan (hakkari) uludağ (Bursa) karagöl, Bolkar mountains (niğde) i n

n = 54 n = 12 n = 18 f r o

min m max sd min m max sd min m max sd g s

o

svl 48.69 57.50 70.68 4.43 51.46 56.73 61.35 2.84 43.03 47.29 52.50 2.56 f

t hl 15.34 17.53 20.19 1.24 14.41 17.26 19.58 1.30 14.17 14.93 16.39 0.69 h e

hW 16.94 19.89 23.67 1.41 17.36 19.86 22.09 1.52 15.66 17.70 19.29 0.97 B e r

id 3.40 4.21 5.10 0.42 3.84 4.31 5.04 0.38 2.86 3.53 4.10 0.33 ç e l io 3.04 3.88 4.72 0.38 3.21 3.71 4.19 0.33 3.03 3.46 3.76 0.23 a n

fl 24.01 28.88 36.20 2.52 24.26 28.45 31.26 2.14 20.68 22.32 24.77 1.26 P l tl 27.87 31.34 39.49 2.57 29.01 31.77 34.46 1.74 20.54 24.01 26.57 1.83 a t e

AAu 27.14 32.93 41.34 2.76 29.12 31.47 35.42 1.98 22.19 25.69 29.87 2.02 a u

(

CAl 2.42 3.01 4.05 0.39 2.12 2.86 3.39 0.35 1.68 2.07 2.49 0.23 h a

hl/svl 0.28 0.31 0.33 0.01 0.28 0.30 0.32 0.01 0.30 0.32 0.34 0.01 k k hW/svl 0.32 0.35 0.38 0.01 0.33 0.35 0.38 0.01 0.35 0.37 0.40 0.013 a r i )

id/svl 0.06 0.07 0.09 0.008 0.07 0.08 0.08 0.004 0.06 0.07 0.08 0.006 ,

e io/svl 0.05 0.07 0.08 0.005 0.06 0.07 0.07 0.004 0.06 0.07 0.08 0.006 a s t

fl/svl 0.44 0.50 0.55 0.02 0.46 0.50 0.53 0.02 0.45 0.47 0.49 0.01 A n

tl/svl 0.49 0.55 0.61 0.02 0.52 0.56 0.59 0.01 0.46 0.51 0.55 0.02 a t o

AAu/svl 0.52 0.57 0.65 0.02 0.50 0.56 0.60 0.03 0.49 0.54 0.57 0.02 l i CAl/svl 0.04 0.05 0.08 0.007 0.04 0.05 0.06 0.005 0.03 0.04 0.05 0.005 a 1 9 Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 6

20 m. A fsAr & B. A fsAr & h. A rikAn

figs. 4a-4b: ventral aspects of a male (a) and female (b) of Rana macrocnemis Boulenger , 1885, from the Berçelan Plateau (hakkari). Abb. 4a-4b: ventralansichten einer männlichen (a) und weiblichen (b) Rana macrocnemis Boulenger , 1885 vom Berçelan Plateau (hakkari).

test. results with p < 0.05 point to a statis - 370 volts for 12 gel columns) until the tically significant difference in the com - tracking dye (bromphenol blue) moved pared characteristics. the morphometric about 53 mm from the start. measurements obtained from the specimens the blood-serum proteins were sepa - of the Berçelan Plateau (hakkari) as well as rated according to the methods given by from the comparative specimens of uludağ dAvis (1964) and slightly modified by (R. macrocnemis ) and mt. Bolkar, karagöl Özeti & A tAtür (1979). Accordingly, a (Rana holtzi ) were subject to independent buffer system containing the stacking gel samples t-test and discriminant function (2.5 %, ph 6.7) at the top and a mixture of Analysis (dfA) using log 10 transformed the separation gel (7.5 %, ph 9.0) and tris- values. glycine (ph 8.3) at the bottom was used. electrophoretic analysis.- By the electrophoretic separations were car - the help of a hematocrit capillary tube, ried out at room temperature (approx. 20-25 blood samples were taken from the cardiac °C). the separation gels were stained with ventricle of adult that had been 0.5 % Amido Black (naphthol Blue Black anesthetized with ether. the blood was cen - 10-B) and then destained passively with trifuged at 600 g for 5 min, separated from repeated 7 % acetic acid baths. later on, the the cellular section, and serum preserved at stained gels were photographed. Qualita - -20 °C. separation was carried out at a lab - tive evaluation of the gels was made direct - oratory temperature of 20 to 25 °C by using ly from the electropherograms, and the den - 5 µl of serum per sample. A Canalco ® sitometric curves of the separations were model 1200 was used as the electrophoresis obtained by means of a gelman ® ACd-15 bath, and a gelman ® deluxe model served model 39430 densitometer at a wavelength as power supply. glass gel columns (4.7 of 500 nm. the images of the stained gels mm x 75 mm) were utilized. Amperage per were taken with a nikon ® Coolpix P1 digi - column was adjusted to 0.85 mA initially tal camera. and gradually increased to 1.75 mA (36 mA, Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 7

the mountain frogs of the Berçelan Plateau (hakkari), east Anatolia 21

results

Color-pattern.- the dorsal skin is = 70, p < 0.05). no differences were found granulated in all hakkari specimens, and the in the measurement ratios between the the head, the back and the dorsal side of the populations of hakkari and uludağ [i.e., R. legs are mostly of light brownish green, macrocnemis ], whereas there were differ - greyish green or yellowish green color. A ences in all ratios obtained except for light brick-red is seen in some specimens id/svl between the populations of hakkari with a light brownish ground color. the and the Bolkar mountains [i.e., R. holtzi ] dorsolateral folds are well developed. there (table 3). in the hakkari population, the is a swelling (mating callus) on the first fin - values for CAl, tl and AAu are bigger ger in all male specimens. the vertebral than in the Bolkar population ( df = 70, p < band is clearly present in 27 specimens (50 0.05, t = 4.88, 5.71 and 3.81, respectively), %) and hardly visible in two specimens whereas those for hW and io are smaller (3.70 %). the population of the Berçelan (df = 70, p < 0.05, t = 8.33 and 3.79, respec - Plateau (hakkari) was divided into four tively). groups according to the presence and discriminant function analysis distin - arrangement of dorsal spots. A large num - guished two distinct groups when the data ber of individual spots of various shapes are of the mountain frogs from hakkari (n = irregularly scattered in 17 of 25 specimens 54), uludağ (n = 12) and the Bolkar lacking a dorsal band, while few spots are mountains (n = 18) was processed. one irregularly scattered in 8 of them (figs. 3a, cluster was formed by the populations of 3b). in 17 of 29 specimens showing a ver - hakkari and uludağ and the other group tebral band, individual spots are scattered in comprised the individuals of the Bolkar an almost symmetrical order to the sides of mountains. the first canonical variate the vertebral band. in 12 specimens, how - explained 92.5 % of the total variance and ever, the spots join to form aggregates on separated the population of the Bolkar the sides of the vertebral band and extend mountains from the other two populations along the paravertebral area (figs. 3c, 3d). (hakkari and uludağ). the first two dis - the spots in the upper part of the hind legs criminant functions explained 100 % of the are arranged in the form of transverse total variance (table 4). in this analysis, bands. the throat region bears sparsely or 100 % of the Bolkar mountains specimens clearly visible dark spots in 30 specimens were classified correctly and formed a clus - but is spotless in 24 specimens. the ventral ter, 11 specimens (20.80 %) of the hakkari side is spotless, off-white or slightly pinkish population were classified incorrectly and gray in males and light pink in females (fig. lumped together with the uludağ speci - 4). the underside of the femur is pinkish or mens, whereas two specimens from the pinkish-gray in males and females. uludağ population (16.70 %) were classi - morphometric measurements fied incorrectly and placed with most and ratios.- the descriptive statistics of hakkari (Berçelan Plateau) specimens the body measurements and ratios of the (table 5 and fig. 5). thereupon, with the populations are provided in table 1. sexual aim to further increase the discriminating dimorphism was expressed by the femur power and improve correct classification, length (fl) in the studied population of the another analysis was made to discriminate Berçelan Plateau (hakkari) ( t = 2.026, df = the populations of the Bolkar mountains 53, p = 0.048) (table 2). independent-sam - from the combined populations “hakkari ples t-tests did not find differences between plus uludağ“. in this analysis, 100 % of the hakkari and uludağ [i.e., R. macrocne - both the Bolkar mountains individuals and mis ] populations, but did so between the the combined “hakkari plus uludağ“ indi - populations of hakkari and the Bolkar viduals were classified correctly (tables 6. mountains [i.e., R. holtzi ] in all the charac - and 7). teristics measured (table 3). the specimens electrophoretic analysis.- sex- in the hakkari population are larger than dependent variation was not observed in the those in the Bolkar population ( t = 9.95, df electropherograms obtained from the blood Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 8

22 m. A fsAr & B. A fsAr & h. A rikAn

fig. 5: graph visualizing the result of a discriminant function Analysis based on eight morphological character - istics of the mountain frog populations of the Berçelan Plateau (hakkari) [circles], uludağ (Bursa) [triangles], and karagöl, Bolkar mountains (niğde) [asterisks]. Abb. 5: graphische darstellung des ergebnisses der diskriminanzanalyse von acht morphologischen merkmalen der Braunfrösche des Berçelan Plateaus (hakkari) [kreise], des uludağ (Bursa) [dreiecke] und von karagöl, Bolkar gebirge (niğde) [sternchen].

plasma samples of the Berçelan Plateau male of the hakkari population (fig. 6) and (hakkari) and sarıalan population (uludağ). the uludağ population (fig. 7), the proteins Based on the gel photograph and densito - can be divided into 13 fractions or fraction metric curve of the electrophoretic separa - groups (one in the albumin region and 12 in tion of the blood-plasma proteins in an adult the region of globulins).

disCussion

Previous studies on the morphology of in the present study, the frogs of the various Anatolian mountain frog popula - Berçelan Plateau (hakkari) were distin - tions ( BArAn 1969; B ArAn & A tAtür 1986; guished easily from R. holtzi by their (i) Çevik et al. 2006; BArAn et al. 2007; granulated skin, (ii) the high rate (53.70 BAşkAle et al. 2012) agree on the observa - %) of a vertebral band present and (iii) tions that in R. holtzi (i) the dorsal skin is the absence of a ring, lighter than the smooth and soft, (ii) no vertebral band is ground color, encircling the spots on the present and (iii) a ring lighter than the dorsum. ground color surrounds each dorsal spot, Rana macrocnemis pseudodalmatina unlike R. macrocnemis . eiselt & s Chmidtler , 1971 was described Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 9

the mountain frogs of the Berçelan Plateau (hakkari), east Anatolia 23

table 2: independent samples t-test compar - table 3: independent samples t-test compar - isons of male and female mountain frogs of the isons of mountain frogs of the Berçelan Plateau (hak- Berçelan Plateau (hakkari), based on measurements of kari) versus specimens of mt. uludağ (Bursa) and ka- nine morphometric variables. the statistically signifi - ragöl, Bolkar mountains (niğde) regarding eight mor - cant ( p < 0.05) difference in femur length (fl) was phometric variables and seven derived ratios. signifi- indicated with an asterisk. see material and methods cant p-values are indicated by an asterisk. see materi- for abbreviations. als and methods for abbreviations in column one. tab. 2: t-test-vergleich männlicher und weib - tab. 3: t-test-vergleich der Braunfrösche des licher Braunfrösche des Berçelan Plateaus (hakkari) Berçelan Plateaus (hakkari) gegenüber fröschen des hinsichtlich neun morphometrischer variablen. der mt. uludağ (Bursa) sowie von karagöl, Bolkar ge- signifikante ( p < 0,05) unterschied in der femurlänge birge (niğde) in acht morphometrischen variablen und (fl) ist mittels Asterisk gekennzeichnet. die sieben abgeleiteten Quotienten. signifikante p-Werte Abkürzungen in spalte eins sind im Abschnitt sind mittels Asterisk gekennzeichnet. die Abkürzun- materials and methods erklärt. gen in spalte eins sind im Abschnitt materials and methods erklärt. Characters males vs. females merkmale männchen gegenüber Weibchen Character Berçelan vs. Berçelan vs. t df p merkmal uludağ karagöl t df p t df p svl 0.29 53 0.766 hl -0.15 53 0.885 svl 0.51 64 0.613 9.95 70 0.000* hW -0.43 53 0.667 hl 0.70 64 0.488 9.01 70 0.000* io 0.91 53 0.366 hW 0.09 64 0.930 6.38 70 0.000* fl 2.03 53 0.048* io 1.36 64 0.180 4.403 70 0.000* tl 1.04 53 0.304 tl -0.63 64 0.529 12.33 70 0.000* AAu 1.76 52 0.085 AAu 1.73 63 0.089 11.15 69 0.000* CAl -0.36 53 0.719 CAl 1.19 64 0.240 10.97 70 0.000* id 0.24 53 0.815 id -0.84 64 0.406 6.56 70 0.000* hW/svl -0.87 64 0.387 -8.33 70 0.000* hl/svl 0.31 64 0.76 -3.19 70 0.002* id/svl -1.19 64 0.23 -0.72 70 0.472 io/svl 1.25 64 0.21 -3.79 70 0.000* tl/svl -1.91 64 0.06 5.71 70 0.000* AAu/svl 1.83 63 0.07 3.81 69 0.000* from the mazandaran province (north iran), CAl/svl 0.96 64 0.34 4.88 70 0.000* a region relatively close to the present study area. in the above study, pseudodalmatina was distinguished from the nominate sub - species by many morphological characteris - tics, including the thin and petite body structure, smoother skin, more barely per - total length (svl) ( BArAn et al. ceptible dorsolateral folds, barely percepti - 2007) and the relative size of the inner ble dorsal pattern, and the absence of a light metatarsal tubercle (CAl) (derived from vertebral band in the middle of the dorsum. data in Çevik et al. 2006) were found to be furthermore, different from R. macrocne - smaller in R. holtzi than in other mountain mis or R. camerani , the white or whitish frog species in Anatolia. BAşkAle et al. upper labial stripe was thinner and more (2012) detected statistically significant dif - petite and extended in cranial direction only ferences in all 12 metric characteristics as far as under the eye. measured when he compared R. holtzi from the mountain frog specimens of the karagöl, Çamlıyayla (mersin) and R. Berçelan Plateau can be distinguished easi - macrocnemis from seviçova (ereğli) and ly from pseudodalmatina by morphological uludağ (Bursa). in the present study, the traits such as the granulated skin, the exten - independent samples t-test detected differ - sion of the white or whitish upper labial ences in eight metric characteristics stripe up to the tip of the chin, the evident between the populations of the Berçelan dorsolateral folds, the distinct dorsal pat - Plateau (hakkari) and mt. Bolkar (karagöl, tern, and the high rate of of a vertebral band niğde) (table 3). the discriminant analy - present. thus, the population of the sis, distinguished clearly the populations of Berçelan Plateau substantially resembles the Berçelan Plateau (hakkari) from the the characteristics given for R. macrocne - population of R. holtzi in the Bolkar mis in the literature. mountains (karagöl). the plot shows a Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 10

24 m. A fsAr & B. A fsAr & h. A rikAn

fig. 6 : gel photograph (below) and densitometric tracing curve (above) showing the electrophoretic separation of the blood plasma proteins obtained from an adult male specimen representing the population of the Berçelan Plateau (hakkari). o.d. - optical density, s - start, A - albumin, g (1-12) - globulins. Abb. 6: gel (unten) und densitometrische verteilungskurve (oben) der elektrophoretischen Auftrennung der Blutplasmaproteine eines adulten männlichen Braunfrosches vom Berçelan Plateau (hakkari). o.d. – optische dichte, s – startpunkt, A Albumin, g (1-12) - globuline.

fig. 7: gel photograph (below) and densitometric tracing curve (above) showing the electrophoretic separation of the blood-plasma proteins obtained from an adult male specimen representing the sarıalan / uludağ (Bursa) population. o.d. - optical density, s - start, A - albumin, g (1-12) - globulins. Abb. 7: gel (unten) und densitometrische verteilungskurve (oben) der elektrophoretischen Auftrennung der Blutplasmaproteine eines adulten männlichen Braunfrosches der sarıalan / uludağ (Bursa) population. o.d. – optische dichte, s – startpunkt, A Albumin, g (1-12) - globuline. Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 11

the mountain frogs of the Berçelan Plateau (hakkari), east Anatolia 25

table 4: descriptive statistics of the first and second canonical discriminant functions of an analysis in which mountain frog populations of the Berçelan Plateau (hakkari), mt. uludağ (Bursa) and karagöl, Bolkar mountains (niğde) were analyzed with regard to eight morphometric variables. tab. 4: deskriptive statistiken der ersten und zweiten kanonischen diskriminanzfunktion aus einer Analyse, in welcher Braunfrösche des Berçelan Plateaus (hakkari), des uludağ (Bursa) und von karagöl, Bolkar gebirge (niğde) hinsichtlich acht morphometrischer variablen untersucht wurden.

Canonical eigenvalue Canonical test of function(s) df significance function Correlation Wilks’ lambda Chi-square 1 4.349 0.902 0.138 154.255 10 0.00* 2 0.351 0.510 0.740 23.453 4 0.00*

table 5: Classification results (predicted group memberships) of the discriminant function analysis of eight morphometric variables from mountain frogs of the Berçelan Plateau (hakkari), mt. uludağ (Bursa) and karagöl, Bolkar mountains (niğde). tab. 5: die zuordnungsergebnisse (vorhergesagte klassenzugehörigkeit) aus der diskriminanzanalyse acht morphologischer variablen von Braunfröschen des Berçelan Plateaus (hakkari), des uludağ (Bursa) und von karagöl, Bolkar gebirge (niğde).

groups / klassen Predicted group membership / vorhergesagte klassenzugehörigkeit ∑ Berçelan uludağ karagöl n Berçelan (hakkari) 42 11 0 53 uludağ (Bursa) 2 10 0 12 karagöl (Bolkar mt.) 0018 18 % Berçelan (hakkari) 79.2 20.8 0.0 100.0 uludağ (Bursa) 16.7 83.3 0.0 100.0 karagöl (Bolkar mt.) 0.0 0.0 100.0 100.0

table 6: descriptive statistics of the first canonical discriminant function of an analysis, in which the com - bined mountain frog populations “Berçelan Plateau (hakkari) plus mt. uludağ (Bursa)” and the individuals from karagöl, Bolkar mountains (niğde) were analyzed with regard to eight morphometric variables. tab. 6: deskriptive statistiken der ersten kanonischen diskriminanzfunktion aus einer Analyse, in welcher die vereinigten Braunfrosch-stichproben “Berçelan Plateau (hakkari) plus uludağ (Bursa)” und die individuen von karagöl, Bolkar gebirge (niğde) hinsichtlich acht morphometrischer variablen untersucht wurden.

Canonical eigenvalue Canonical test of function(s) df significance function Correlation Wilks’ lambda Chi-square 1 4.301 0.901 0.189 130.936 5 0.00*

table 7: Classification results (predicted group memberships) of the discriminant function analysis of eight morphometric variables from the combined mountain frog populations “Berçelan Plateau (hakkari) plus mt. uludağ (Bursa)” and the individuals from karagöl, Bolkar mountains (niğde). one hundred percent of the cases were classified correctly. tab. 7: die zuordnungsergebnisse (vorhergesagte klassenzugehörigkeit) aus der diskriminanzanalyse acht morphologischer variablen von Braunfröschen der vereinigten stichproben “Berçelan Plateau (hakkari) plus uludağ (Bursa)” und der individuen von karagöl, Bolkar gebirge (niğde).

groups / klassen Predicted group membership / vorhergesagte klassenzugehörigkeit ∑ Berçelan + uludağ karagöl n Berçelan + uludağ 65 0 65 karagöl (Bolkar mt.) 0 18 18 % Berçelan + uludağ 100.0 0.0 100.0 karagöl 0.0 100.0 100.0 Afsar_etal_mountainfrogs_from_hakkari_tr_herPetozoA.qxd 28.07.2015 14:54 seite 12

26 m. A fsAr & B. A fsAr & h. A rikAn

distinct separation between the populations teins, while only genetic variations cause of the Berçelan Plateau and the Bolkar qualitative differences. interpopulational mountains (karagöl); however, the popula - quantitative differences were in fact evi - tion of the Berçelan Plateau cannot be dis - dent, especially among the globulins, but tinguished from the population of R. macro- are not of taxonomical significance. this cnemis from sarıalan (uludağ) (figure 5 result is in agreement with the result and table 5). provided for the uludağ mountain frogs ( R. As regards the blood-plasma proteins, macrocnemis ) by Çevik et al. (2006). significant quantitative differences are pres - in conclusion, the population of the ent within the regions of globulins of the Berçelan Plateau (hakkari) substantially population belonging to the Berçelan Pla - resembles the sarıalan population (uludağ) teau (hakkari) and the sarıalan population when both morphological characteristics (uludağ). According to dessAuer (1956), and the results of the electrophoretic analy - Chen (1967), ferguson (1980) and ArikAn sis of the blood-plasma proteins are taken et al. (1998, 1999), who worked on the into consideration. hence, the authors are blood-plasma proteins of , fac - convinced that it is appropriate to assign this tors such as age, as well as physiological east Anatolian mountain frog population to and environmental conditions, cause quanti - the taxon R. macrocnemis . tative differences in the blood-plasma pro -

ACknoWledgments

this study was financially supported within Affairs ministry of nature Protection and national Project no. fef 2010-062 by the scientific research Parks and research permission was given by decision Projects at Celal Bayar university. the working pro - no. 2011/ B.18.0.dmP.0.02-510.02-47728. cedures were approved by the forestry and Water

referenCes

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dAte of suBmission: January 16, 2014 Corresponding editor: heinz grillitsch

Authors: murat AfsAr (Corresponding author < [email protected] >) , Birgül AfsAr – department of Biology, faculty of Arts & sciences, Celal Bayar university, muradiye Campus, 45030, manisa, turkey; hüseyin ArikAn – department of Biology, faculty of science, ege university, 35100, Bornova, İzmir, turkey.